Candida is a fungus that is among the most common causes of infections in hospitalized patients in the United States and worldwide. Candida spp. are 1 of the most common microbes that infect the blood and urine of hospitalized patients. The cost associated with Candida blood infections alone exceeds at least $1 billion per year in the US. Even with therapy, 40-50% of patients with Candida blood infections die from the infection. Furthermore, resistance to antifungal therapies is rising among Candida organisms. For these reasons, a vaccine to prevent life threatening Candida infections is particularly attractive. Such a vaccine could potentially be useful for large populations including nearly all patients who undergo surgery to the chest or abdomen, newborns in intensive care units, and other patients with weakened immune systems, including cancer patients receiving chemotherapy. We have isolated a protein that allows Candida to adhere to human cells. The gene that encodes this protein is called ALS1, and it is a member of a family of related genes that code for at least 9 proteins, most of which are also adhesins. Active immunization with the recombinant N-terminus of Als1p (rAls1p-N) significantly protects mice from otherwise rapidly lethal bloodstream and deep organ Candida infection. More recently we have purified a related protein, rAls3p-N, that is also encoded by a member of the ALS gene family. We have discovered that rAls3p-N results in a broader immune response than rAls1p-N, raising the possibility that rAls3p-N might be more effective than rAls1p-N as a vaccine. We propose to define and optimize vaccine-mediated protection in preparation for future clinical studies in humans by identifying the optimally protective Als protein (Alsp) in murine models of infection. The efficacy of rAls1p-N vs. rAls3p-N vs. combination rAls1p-N + rAls3p-N vs. adjuvant alone will be compared in our murine models of bloodstream infection, oral thrush, or vaginal yeast infection. Although the current application is focused on the development of an anti-Candida vaccine, we have extremely novel data showing cross-protection of the rAls1p-N vaccine against S. aureus in the murine model. Therefore, we will also compare the efficacy of the vaccine immunogens in our murine model of S. aureus bloodstream infection. Finally, in other systems, activation of a host immune receptor called TLR5 has been shown to enhance the efficacy of vaccines. Therefore, to further enhance the immune response to the Alsp immunogens, we will create novel combination proteins by mixing the Alsp immunogens with special proteins that activate TLR5. Once we have optimized our immunogens, we intended to prepare the vaccine product for human trials in the future phases of this project. Due to modern therapeutic advances for sustained life support, patients undergoing a variety of medical procedures or with a variety of medical conditions have become at risk for developing life-threatening infections caused by the fungus Candida. Among these procedures and conditions are: being treated in an intensive care unit, having a plastic catheter in a large vein, undergoing surgery to the chest or abdomen, being the victim of trauma injuries (both domestic, such as motor vehicle accidents, and military), having extensive burn injuries, being a premature neonate, and having cancer and undergoing chemotherapy. Given how common these conditions are, the development of a vaccine that protects against Candida infections could save hundreds of thousands of lives as well as substantially reduce hospitalization costs in both the United States and other countries where highly effective medical advances have been made. [unreadable] [unreadable] [unreadable]